CO2 Assimilation and Water Relations of Almond Tree (Prunus amygdalus Batsch) Cultivars Grown Under Field Conditions

Gas exchanges and leaf water potential (_w) of six-years-old trees of fourteen Prunus amygdalus cultivars, grafted on GF-677, were studied in May, when fruits were in active growing period, and in October, after harvesting. The trees were grown in the field under rain fed conditions. Predawn _w showed lower water availability in October compared with May. The lowest _w values at midday in May increased gradually afterwards, while in October they decreased progressively until night, suggesting a higher difficulty to compensate the water lost by transpiration. However, relative water content (RWC) measured in the morning was similar in both periods, most likely due to some rainfall that occurred in September and first days of October that could be enough to re-hydrate canopy without significantly increasing soil water availability. The highest net photosynthetic rate (P _N) was found in both periods early in the morning (08:00–11:00). Reductions in P _N from May to October occurred in most cultivars except in José Dias and Ferrastar. In all cultivars a decrease in stomatal conductance (g _s) was observed. Photosynthetic capacity (P _max) did not significantly change from spring to autumn in nine cultivars, revealing a high resistance of photosynthetic machinery of this species to environmental stresses, namely high temperature and drought. Osmotic adjustment was observed in some cultivars, which showed reductions of ca. 23 % (Duro d' Estrada, José Dias) and 15 % (Tuono) in leaf osmotic potential (). Such decreases were accompanied by soluble sugars accumulation. The Portuguese cultivar José Dias had a higher photosynthetic performance than the remaining genotypes.     

Proline accumulation and nitrate reductase activity in contrasting sorghum lines during mid-season drought stress

Six lines of sorghum ( Sorghum bicolor L. Moench) with differing drought resistance (IS 22380, ICSV 213, IS 13441 and SPH 263, resistant and IS 12739 and IS 12744, susceptible) were grown under field conditions in the semi-arid tropics and analysed for proline and nitrate reductase activity (NRA; EC 1.6.6.1) during a mid-season drought. The resistant lines accumulated high levels of proline, while the susceptible lines showed no significant proline accumulation. Most of the proline was accumulated after growth of the plants had ceased. In a separate greenhouse experiment, most of the proline was found in the green rather than the fired portions of leaves. The levels returned to that of irrigated controls within 5 days of rewatering. Proline levels increased as leaf water potential and relative water content fell, and there was no apparent difference among the different sorghum lines with change in plant water status. Susceptible lines accumulated less proline than resistant lines as leaf death occurred at higher water potentials. Proline accumulation may, however, contribute to the immediate recovery of plants from drought. Leaf NRA reached high levels at about 35 days after sowing in both the stressed and irrigated plants, after which it declined. The decline in NRA was more pronounced in the stressed than in the irrigated plants and closely followed changes in the growth rate. Upon rewatering, NRA increased several-fold in all the lines and, in contrast to proline accumulation, genotypic differences in NRA were small, both during stress and upon rewatering. The high sensitivity of NRA to mild drought stress was reflected in the rapid decline of activity with small changes in leaf water potential and relative water content. The results are discussed in the light of a possible role for proline during recovery from drought, and the maintenance of NRA during stress and its recovery upon rewatering.     

Role of proline and leaf expansion rate in the recovery of stressed white clover leaves with increased phosphorus concentration

Osmotic adjustment (OA) and increased cell-wall extensibility required for expansive leaf growth are well defined components of adaptation to water stress in dry soil, which might interact with soil phosphorus (P) concentration and defoliation frequency for intensively grazed white clover in legume-based pastures. Experiments were conducted with frequently and infrequently defoliated mini-swards of white clover growing in dry soil with low and high P concentrations. The higher yielding high-P plants were able to dry the soil to greater soil water suctions; their leaves had lower water potential values, yet they showed fewer water stress symptoms and underwent a more complete recovery from the water stress symptoms on rewatering, than the low-P plants. High- P plants had greater OA, proline concentration and leaf expansion rate. On the other hand, low-P plants showed an increased osmotic concentration when there was no change in the total solute content per unit of leaf d. wt, indicating more loss of water from the leaf tissue. The key measures that appeared to be directly associated with plant recovery over a short period following water stress were increased proline concentration and leaf expansion rate, probably resulting from increased cell-wall extensibility rather than increased production of cells for the high-P plants.     

Phosphoenolpynivate carboxylase, malate and alcohol dehydrogenase activities in alfalfa (Medicago sativa) nodules under water stress

The effect of drought upon phosphoenolpyruvate carboxylase (PEPC; EC 4.1.1.31), malate ddiydrogenase (MDH; EC 1.1.1.37), alcohol dehydrogenase (ADH; EC 1.1.1.1) and β -hydroxybulyrate dehydrogenase ( β -OH-BDH; EC 1.1.1.30) enzyme activities as well as the leghemoglobin (Lb), malate and ethanol contents of alfalfa nodules ( Medicago sativa L. cv. Aragon) were examined. Both the ieghemoglobin (Lb) content and the Lb/soluble protein ratio were significantly reduced at a nodule water potential (Ψ_nod) of—1.3 MPa. At lower Ψ_nod, Lb content decreased further, but the ratio remained unchanged. Slight stress (—1.3 MPa) drastically affected acetylene reduction activity (ARA; 60% reduction) whereas in vitro PEPC activity was main-tained at relatively constant values. As stress progressed (—2.0 MPa), a simultaneous reduction in both activities was observed. Severe stress (Ψ_nod lower than —2.0 MPa) stimulated in vitro PEPC. Bacteroid β -J-OH-BDH activity was stimulated by slight (—1.3 MPa) and moderate (—2.0 MPa) drought. MDH activity rose in slightly stressed nodules (Ψ_nod—1.3 MPa). Greater water deficits sharply decreased MDH activity to values significantly lower than those found in control nodules. Nodule malate content followed the same pattern as MDH. The plant fraction of the nodule showed constitutive ADH activity and contained ethanol. ADH was stimulated at slight (— 1.3 MPa) and moderate drought levels (—2.0 MPa). Ethanol content showed similar behavior to ADH activity. Inhibition of ARA, reduction of Lb content and stimulation of the fermentative metabolism induced by water stress suggest some reduction ira O₂ availability within the nodule.     

Growth, Water Content, and Proline Accumulation in Drought-Stressed Callus of Date Palm

This study was conducted to examine the response of date palm (Phoenix dactylifera L., cvs. Barhee and Hillali) calli to water stress. Callus derived from shoot tip explants was inoculated in liquid Murashige and Skoog medium containing 10 mg dm^–3-naphthaleneacetic acid, 1.5 mg dm^–3 2-isopentenyladenine, and 0 to 30 % (m/v) polyethylene glycol (PEG 8000) to examine the effect of water stress. After 2 weeks, callus growth, water content, and proline accumulation were assessed. Increasing water stress caused a progressive reduction in growth as expressed in callus fresh mass, relative growth rate, and index of tolerance. Both genotypes tested followed this general trend, however, cv. Barhee was more tolerant to drought stress than cv. Hillali. Increasing PEG concentration was also associated with a progressive reduction in water content and increased content of endogenous free proline.     

Osmotic adjustment inSorghum bicolor (L. Moench) grown under moisture stress in soil and osmotically modified solution cultures

Sorghum (Sorghum bicolor L. Moench) plants were grown in solution culture and stressed at three rates of decreasing leaf water potential (−0.123, −0.068 and −0.029 MPa day⁻¹) achieved by the incremental addition of an osmoticum, polyethylene glycol (PEG) 6000 to the solutions. Plants were also grown in soil and given different amounts of water which resulted in rates of decreasing leaf water potentials of −0.130 and −0.073 MPa day⁻¹. The rate of stress and the culture system influenced the accumulation of solutes in the cell, but not cell volume. A rapid stress (−0.123 and −0.130 MPa day⁻¹) to approximately −1.6 MPa leaf water potential resulted in 0.75 and 0.16 MPa of osmotic adjustment in the PEG and soil culture respectively. At moderate stress (−0.068 and −0.073 MPa day⁻¹) respective values were 1.68 and 0.58 MPa. There were some visual symptoms in the solution grown plants characteristic of uptake of high molecular weight PEG. However the relative growth rates of these plants were equal to or greater than those of the soil grown plants. In view of the differences in plant water status of soil and PEG solution cultured plants it was concluded that the use of the latter system would not be entirely suitable for some studies of drought resistance in sorghum, as related to crop performance in the field.     

In vitro-selection of hydroxyproline-resistant cell lines of wheat (Triticum aestivum): accumulation of proline, decrease in osmotic potential, and increase in frost tolerance

Five hundred hydroxyproline-resistant cell lines were selected from cell cultures of wheat ( Triticum aestivum L. cv. Koga II) after plating on 10 to 30 m M hydroxyproline (Hyp) containing solid Gamborg B 5 medium. All selected cell lines from 30 m M Hyp-medium contained increased (up to 17-fold) levels of free proline. Seventy-four cell lines were transferred to Hyp-free medium and subcultivated 25 times, for 12 months altogether, after which 80% still had increased proline levels. Fourteen cell lines with increased proline levels were further investigated in liquid media with regard to their frost tolerance, which was measured by means of electrolyte leakage. Ten of them showed increased fros tolerance, with LT 50 values as low as 2.7°C below that of the wild type (-4.7°C). Besides increased proline levels and increased percentage dry weight, the Hyp-resistant cell lines had lower osmotic potentials. Osmotic potentials correlated better than levels of free proline with the increase in frost tolerance.     

Drought tolerance in faster- and slower-growing black spruce (Picea mariana) progenies: II. Osmotic adjustment and changes of soluble carbohydrates and amino acids under osmotic stress

The possibility was considered that osmotic adjustment, the ability to accumulate solutes in response to water stress, may contribute to growth rate differences among closely-related genotypes of trees. Progeny variation in osmotic adjustment and turgor regulation was investigated by comparing changes in osmotic and pressure potentials, soluble carbohydrates, and amino acids in osmotically stressed seedlings in 4 full-sib progenies of black spruce [ Picea mariana (Mill.) B. S. P.] that differed in growth rate under drought. Osmotic stress was induced by a stepwise increase in the concentration of polyethylene glycol (PEG)-3350 from 10 (w/v) to 18 and 25%, which provided osmotic potentials in solution culture of -0.4, -1.0 and -2.0 MPa each for 3 days. All 4 progenies maintained a positive cell turgor even at 25% PEG, due to a significant decline in osmotic potential. Although total amino acids, principally proline, increased, ca 60% of the decrease in osmotic potential was attributable to soluble carbohydrates and glucose was the major osmoregulating solute. There was little progeny variation in any of measured parameters in unstressed seedlings. Compared to two slower-growing progenies, the two progenies capable of more vigorous growth under drought in the field accumulated more soluble carbohydrates (mainly glucose and fructose), developed lower osmotic potential and maintained higher turgor pressure when osmotically-stressed in solution culture. The ability to adjust osmotically and maintain turgor under drought stress could thus be a useful criterion for the early selection of faster-growing, drought-tolerant genotypes.     

Water stress induced alterations in ornithine aminotransferase of ragi (Eleusine coracana): Protection by proline against heat inactivation and denaturation by urea and guanidinium chloride

Water stress resulted in a specific response leading to a large and significant increase (80-fold) in free proline content of ragi (Eleusine coracana leaves and seedlings. L-Proline protected ornithine aminotransferase, an enzyme in the pathway for proline biosynthesis, isolated from normal and stressed ragi leaves against heat inactivation and denaturation by urea and guanidinium chloride. The protection of the stressed enzyme by L-proline was much more complete than that of the enzyme isolated from normal leaves. While L-ornithine, one of the substrates, protected the stressed enzyme against inactivation, it enhanced the rate of inactivation of the normal enzyme. α-Ketoglutarate protected both the normal and stressed enzyme against inactivation and denaturation. These results support the suggestion that ornithine aminotransferase has undergone a structural alteration during water stress. In view of the causal relationship between elevated temperature and water stress of plants under natural conditions, the protection afforded by proline against inactivation and denaturation of the enzyme from stressed leaves assumes significance. These results provide an explanation for a possible functional importance of proline accumulation during water stress.     

Influence of Phosphorus Application on Water Relations, Biochemical Parameters and Gum Content in Cluster Bean Under Water Deficit

Relative water content (RWC), leaf water potential (_w) and osmotic potential (_s), contents of chlorophyll (Chl) a, Chl b, soluble sugars, and seed quality (gum content) were used to evaluate the role of phosphorus in alleviation of the deleterious effect of water deficit in clusterbean (Cyamopsis tetragonoloba L. Taub). Under water stress, _w, _s, and Chl and gum contents decreased and soluble sugar contents increased. Phosphorus application increased Chl and sugar contents in control plants and ameliorated negative effects of water stress.     

The pattern of distribution of pectin, peroxidase and lignin in the middle lamella of secondary xylem fibres in alfalfa (Medicago sativa)

BACKGROUNDS AND AIMS: Information on the micro-distribution of lignin within the middle lamella is only just beginning to emerge. This paper provides evidence of marked heterogeneity in the micro-distribution of lignin, pectin, peroxidase and hydrogen peroxide in the middle lamella of alfalfa (Medicago sativa). METHODS: Specimens from alfalfa stems were collected and processed for transmission electron microscopy. The middle lamella architecture was examined prior to and during lignification, using transmission electron microscopy in combination with pectin- and lignin-specific staining. In addition, immuno-gold labelling of peroxidase and cytochemical localization of hydrogen peroxide (H2O2) were undertaken. KEY RESULTS: Lignin showed inhomogeneity in its distribution in the middle lamella. It was found that the distribution of pectin was irregular and corresponded to the pattern of deposited lignin. Additionally, a similarity in the pattern of the deposited lignin to the pattern of distribution of peroxidase and H2O2 was also observed. CONCLUSIONS: Irregular distribution of pectin in the middle lamella may be related to subsequent inhomegeneity in lignin in this region.     

Implications of water stress-induced changes in the levels of endogenous ascorbic acid and hydrogen peroxide in Vigna seedlings

Vigna cutjang Endl. cv. Pusa Barsati seedlings, subjected to increasing degrees of water stress (−0.5, −1.0, −1,5 MPa), produced an approximately proportional increase in glycolate oxidase activity, hydrogen peroxide (H₂O₂) and proline content but a decrease in catalase activity, ascorbic acid and protein content. Leaf water potential (leaf ψ) and relative water content (RWC) were also lowered with increasing stress. Pretreatment with l -cysteine and reduced glutathione (10-3 M) decreased glycolate oxidase activity, H₂O₂ content, ascorbic acid oxidase activity, proline content and also slightly improved the water status of leaves stressed (−1.0 MPa) for 2 days. Pretreatment of non-stressed seedlings with these antioxidants had little or no effect. These studies indicate that treatment with antioxidants makes the plant tolerant against water stress by modulating the endogenous levels of H₂O₂ and ascorbic acid in stressed tissue.     

Effect of mannitol and glucose-induced osmotic stress on growth,water relations,and solute composition of cell suspension cultures of poplar (Populus deltoides var.Occidentalis) in relation to anthocyanin accumulation

Summary A cell suspension culture of poplar (Populus deltoides (Marsh.) Bartr. var.occidentalis Rydb.), accumulating the anthocyanin pigment, cyanidin 3-glucoside, in the lag phase of culture growth, was subjected to osmotic stress with glucose and mannitol. Osmotic stress treatments resulted in growth suppression and higher anthocyanin accumulation compared with unstressed cells. Both an increase in the proportion of pigmented cells and an increase in the concentration of anthocyanin in the pigmented cells were responsible for high anthocyanin content of cultured cells subjected to osmotic stress. The osmotic stress induced by glucose suppressed growth more than that by mannitol and produced higher anthocyanin levels. Only small amounts of [U-¹⁴C]mannitol were taken up and metabolized by the cells. Stressed cells accumulated sugars and free amino acids to a different extent resulting in altered cell sugar-to-amino acid ratios. The accumulation of osmotically active solutes and cell growth suppression may both be responsible for the accumulation of anthocyanin in stressed cells.     

Effect of water deficit on photosynthetic and other physiological responses in grapevine (Vitis vinifera L. cv. Riesling) plants

The grapevine (Vitis vinifera L. cv. Riesling) plants subjected to water deficit were studied for changes in relative water content (RWC), leaf dry mass, contents of chlorophyll (Chl), total leaf proteins, free amino acids, and proline, and activities of ribulose-1,5-bisphosphate carboxylase (RuBPC), nitrate reductase (NR), and protease. In water-stressed plants RWC, leaf dry matter, Chl content, net photosynthetic rate (P _N), and RuBPC and NR activities were significantly decreased. The total leaf protein content also declined with increase in the accumulation of free amino acids. Concurrently, the protease activity in the tissues was also increased. A significant two-fold increase in proline content was recorded.     

Strain Variation in Tolerance of Water Stress by Idriella (Microdochium) bolleyi,a Biocontrol Agent of Cereal Root and Stem Base Pathogens

Three randomly chosen isolates of Idriella bolleyi differed markedly in tolerance of water stress down to—5 MPa in vitro. The differences were seen with respect to osmotic potential of media adjusted with KCl or matric potential of media adjusted with polyethylene glycol 8000. They were consistent when assessed by linear extension of colonies, spore germination, biomass production in liquid culture and sporulation in liquid culture. In comparative tests, one strain of I. bolleyi showed similar stress tolerance to that oF Fusarium culmorum, noted as a pathogen of cereals in dry conditions, whereas another strain of I. bolleyi showed low tolerance equivalent to that of the take‐all fungus Gaeumannomyces graminis and a Phialophora sp. from maize. The extreme variation in water stress tolerance of I. bolleyi might be used to select strains for biocontrol of different cereal root‐ and foot‐rot pathogens or strains might be combined in seed‐applied inocula for consistency of biocontrol in different site or seasonal conditions.     

Alterations in the biosynthesis of proteins and nucleic acids in finger millet (Eleucine coracana) seedlings during water stress and the effect of proline on protein biosynthesis

The biosynthesis of DNA appeared to be unaffected in the water-stressed seedlings of finger millet (Eleucine coracana), but an increase in the synthesis de novo of RNA and proteins was observed during mild water stress. The polyribosome content was also increased in stressed finger millet seedlings. Proline, a solute which accumulates during water stress, enhanced the incorporation of radioactive precursors into proteins; caused an increase in translatability of finger millet messengers in vitro; and stabilized the polyribosomes isolated from normal seedlings. The results emphasize the role of proline in the adaptation of finger millet to the intermittent drought it experiences during cultivation.